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Key message A total of 45 aquaporins was identified in Fagus sylvatica, 35 of which were differentially expressed in spring and summer in the leaves, phloem and xylem of 11-year-old trees. Abstract European beech (Fagus sylvatica) has been widely studied in terms of its water relations and local adaptation. However, to date, the underlying basis conferring adaptation to differences in water availability are unknown. Therefore, we examined the expression of aquaporins in trees of four different beech provenances representing the southern and northern range margins, as well as core populations, grown in a common garden. We sampled their xylem, phloem and leaf tissue, when leaves had fully expanded, and in late summer. A total of 45 aquaporin isoforms were identified in the beech genome, of which 35 were detected across all sampled tissues. In our phylogenetic analysis, beech aquaporins clustered into the five subfamilies found in other woody species. Members of the plasma membrane intrinsic protein subfamily generally displayed the highest levels of expression, followed by tonoplast intrinsic proteins. Isoforms of the remaining subfamilies, Noduline-26-like intrinsic proteins, small basic intrinsic proteins and uncharacterised intrinsic proteins, were expressed at very low to moderate levels. The expression of most isoforms was stable or declined from spring to summer. Leaves followed a different expression profile from that of vascular tissues, whereas both phloem and xylem were found to express the same FsMIPs. Tissue-specific aquaporin expression was very similar amongst the four beech provenances, indicating that there is no inherent difference in the capability of these provenances to regulate aquaporin activity. The general decrease in FsMIP expression toward the end of the growing period indicates that aquaporins are involved in tree water relations and growth.

Incomplete knowledge of carbon (C) allocation dynamics in trees hinders accurate modeling and future predictions of tree growth. We studied C allocation dynamics in a mature Pinus sylvestris L. dominated forest with a novel analytical approach, allowing the first comparison of: (i) magnitude and delta C-13 of shoot, stem and soil CO2 fluxes (A(shoot), R-stem and R-soil), (ii) concentration and delta C-13 of compound-specific and/or bulk non-structural carbohydrates (NSCs) in phloem and roots and (iii) growth of stem and fine roots. Results showed a significant effect of phloem NSC concentrations on tracheid growth, and both variables significantly impacted R-stem. Also, concentrations of root NSCs, especially starch, had a significant effect on fine root growth, although no effect of root NSC concentrations or root growth was detected on R-soil. Time series analysis between delta C-13 of A(shoot) and delta C-13 of R-stem or delta C-13 of R-soil revealed strengthened C allocation to stem or roots under high C demands. Furthermore, we detected a significant correlation between delta C-13 of R-stem and delta C-13 of phloem sucrose and glucose, but not for starch or water-soluble carbohydrates. Our results indicate the need to include C allocation dynamics into tree growth models. We recommend using compound-specific concentration and delta C-13 analysis to reveal C allocation processes that may not be detected by the conventional approach that utilizes bulk organic matter.

Sunflecks are transient patches of direct radiation that provide a substantial proportion of the daily irradiance to leaves in the lower canopy. In this position, faster photosynthetic induction would allow for higher sunfleck-use efficiency, as is commonly reported in the literature. Yet, when sunflecks are too few and far between, it may be more beneficial for shade leaves to prioritize efficient photosynthesis under shade. We investigated the temporal dynamics of photosynthetic induction, recovery under shade, and stomatal movement during a sunfleck, in sun and shade leaves of Fagus sylvatica from three provenances of contrasting origin. We found that shade leaves complete full induction in a shorter time than sun leaves, but that sun leaves respond faster than shade leaves due to their much larger amplitude of induction. The core-range provenance achieved faster stomatal opening in shade leaves, which may allow for better sunfleck-use efficiency in denser canopies and lower canopy positions. Our findings represent a paradigm shift for future research into light fluctuations in canopies, drawing attention to the ubiquitous importance of sunflecks for photosynthesis, not only in lower-canopy leaves where shade is prevalent, but particularly in the upper canopy where longer sunflecks are more common due to canopy openness.

Xylem sap exists in a state some have described as “doubly” metastable[1]: liquid water is transported from root to leaf under negative pressure, and, in some climates, below its freezing point. Sub-100 nm nanobubbles may be injected into the xylem liquid through pit membranes[2], becoming coated with Phospho- and Glycolipids in the process. Their surface properties, and therefore fate within the tree hydraulic system, remain largely unexplored.In this work, we have used molecular dynamics simulations to produce surface tension – area isotherms of biologically relevant lipid monolayers, as a function of both temperature and negative pressure (i.e. dynamic surface tensions).We find that glycolipid monolayers resist expansion proportionally to the rate of expansion[3]. Their surface tension increases with the tension applied, stabilising the bubble with respect to embolism. In contrast, a typical phospholipid rapidly condenses into more dense lamellar-like phase, rendering it highly resistant to tensions as high as -3.5 MPa. Mixed monolayers of the two exhibit hybrid behavior, as the glycolipids' larger head group disrupts the more ordered phase of the phospholipid. Finally, it is observed that increasing temperature also increases surface tension, at a given surface area.[1] Lintunen, A., Hölttä, T., & Kulmala, M. (2013). Anatomical regulation of ice nucleation and cavitation helps trees to survive freezing and drought stress. Scientific Reports, 3, 1–7. https://doi.org/10.1038/srep02031[2] Schenk, H. J., Steppe, K., & Jansen, S. (2015). Nanobubbles: a new paradigm for air-seeding in xylem. Trends in Plant Science, 20(4), 199–205. https://doi.org/10.1016/j.tplants.2015.01.008[3] Ingram, S., Salmon, Y., Lintunen, A., Hölttä, T., Vesala, T., & Vehkamäki, H. (2021). Dynamic Surface Tension Enhances the Stability of Nanobubbles in Xylem Sap. Frontiers in Plant Science, 12. https://doi.org/10.3389/fpls.2021.7327

The stomata on the leaves of terrestrial plants are a crucial pathway both in the soil-plant-atmosphere hydrological continuum and in the global carbon cycle. Stomatal optimization approaches have proven to be relevant in modelling the trade-off between carbon assimilation and water stress avoidance. In this in-depth case study, we use new optimization-based stomatal models in modelling vegetation gas exchange with the land surface model JSBACH.The theoretical framework presented in Dewar et al. (2018) combines different optimization hypotheses and photosynthesis models to provide analytical solutions for various leaf-level state variables such as stomatal conductance and photosynthesis rate. The most successful combinations assume that plants regulate stomata as if to maximize photosynthesis at all times, and that photosynthesis is restricted by non-stomatal limitations related to water stress. In this study, we further develop the framework, which yields several promising stomatal conductance models.We implement these stomatal models in the land surface model JSBACH, which we run for a single boreal forest site, the SMEAR II measurement station in southern Finland. The model runs are constrained with meteorological and soil moisture data and parametrized with plant properties previously measured at the site, such as xylem hydraulic conductance and photosynthetic parameters. Gross primary production and transpiration rates predicted by JSBACH under different stomatal and photosynthesis models are compared to eddy covariance measurements from SMEAR II, covering the years 2006 through 2012. The model results are also compared to each other and to those obtained using the Unified Stomatal Optimization model by Medlyn et al. (2011). The comparison is restricted to dry daytime hours in the growing season. References:Dewar et al. 2018, New Phytol. 217: 571–581Medlyn et al. 2011, Glob. Change Biol. 17: 2134–2144

Urban vegetation plays an important role in offsetting urban CO2 emissions and mitigating heat through tree transpiration and shading. With frequent heatwave events and the accompanying drought, the functioning of urban trees is severely affected in terms of photosynthesis and transpiration rate. The detailed response is however still unknown despite tree functioning having crucial effects on the ecosystem services they provide. We conducted sap flux density (Js) and leaf gas exchange measurements of trees (Tilia cordata, Tilia × europaea, Betula pendula, Malus spp.) located at four types of urban green areas (Park, Street, Forest, Orchard) in Helsinki, Finland, over two contrasting summers 2020 and 2021. Summer 2021 had a strong heatwave and drought, whereas summer 2020 was more typical for Helsinki. In this study, our aim was to understand the responses of urban tree transpiration and leaf gas exchange to heatwave and drought and examine the main environmental drivers controlling the transpiration rate during these periods in urban green areas. We observed varying responses of tree water use during the heatwave period at the four urban sites. Js was found to be 35–67 % higher during the heatwave as compared to the non-heatwave period at the Park, Forest, and Orchard sites but no significant difference was found at the Street site. Our results showed that Js was higher (31–63 %) at all sites during drought as compared to non-dry periods. The higher Js during the heatwave and dry periods were mainly driven by the high atmospheric demand for evapotranspiration represented by the vapor pressure deficit (VPD), suggesting that the trees were not experiencing severe enough heat or drought stress that stomatal control would have decreased transpiration. Accordingly, maximum assimilation (Amax), stomatal conductance (gs), and transpiration (E) at the leaf level did not change at the four sites during heatwave and drought periods. However, gs was substantially reduced during the drought period at the Park site. VPD explained 55–69 % variations in the daily mean Js during heatwave and drought periods at all sites except at the Forest site where the saturation of Js at high VPD was evident due to low soil water availability. The heat and drought conditions were untypically harsh for the region but not excessive enough to restrict stomatal control and the increased transpiration indicating that ecosystem services such as cooling was not at risk.

In many regions, drought is suspected to be a cause of Scots pine decline and mortality, but the underlying physiological mechanisms remain unclear. Because of their relationship to ecohydrological processes, δ18O values in tree rings are potentially useful for deciphering long-term physiological responses and tree adaptation to increasing drought. We therefore analyzed both needle- and stem-level isotope fractionations in mature trees exposed to varying water supply. In a first experiment, we investigated seasonal δ18O variations in soil and needle water of Scots pine in a dry inner Alpine valley in Switzerland, comparing drought-stressed trees with trees that were irrigated for more than 10 years. In a second experiment, we analyzed twentieth-century δ18O variations in tree rings of the same forest, including a group of trees that had recently died. We observed less 18O enrichment in needle water of drought-stressed compared with irrigated trees. We applied different isotope fractionation models to explain these results, including the Péclet and the two-pool correction, which considers the ratio of unenriched xylem water in the needles to total needle water. Based on anatomical measurements, we found this ratio to be unchanged in drought-stressed needles, although they were shorter. The observed lower 18O enrichment in needles of stressed trees was therefore likely caused by increased effective path length for water movement within the leaf lamina. In the tree-ring study, we observed lower δ18O values in tree rings of dead trees compared with survivors during several decades prior to their death. These lower values in declining trees are consistent with the lower needle water 18O enrichment observed for drought-stressed compared with irrigated trees, suggesting that this needle-level signal is reflected in the tree rings, although changes in rooting depth could also play a role. Our study demonstrates that long-term effects of drought are reflected in the tree-ring δ18O values, which helps to provide a better understanding of past tree physiological changes of Scots pine.

Tree stems are an overlooked source of volatile organic compounds (VOCs). Their contribution to ecosystem processes and total VOC fluxes is not well studied, and assessing it requires better understanding of stem emission dynamics and their driving processes. To gain more mechanistic insight into stem emission patterns, we measured monoterpene, methanol, and acetaldehyde emissions from the stems of mature Scots pines (Pinus sylvestris L.) in a boreal forest over three summers. We analysed the effects of temperature, soil water content, tree water status, transpiration, and growth on the VOC emissions, and used generalized linear models to test their relative importance in explaining the emissions. We show that Scots pine stems are considerable sources of monoterpenes, methanol, and acetaldehyde, and their emissions are strongly regulated by temperature. However, even small changes in water availability affected the emission potentials: increased soil water content increased the monoterpene emissions within a day, whereas acetaldehyde and methanol emissions responded within two to four days. This lag corresponded to their transport time in the xylem sap from the roots to the stem. Moreover, the emissions of monoterpenes, methanol, and acetaldehyde were influenced by the cambial growth rate of the stem with six- to ten-day lags. This article is protected by copyright. All rights reserved.

Photosynthetic rate is concurrently limited by stomatal limitations and nonstomatal limitations (NSLs). However, the controls on NSLs to photosynthesis and their coordination with stomatal control on different timescales remain poorly understood. According to a recent optimization hypothesis, NSLs depend on leaf osmotic or water status and are coordinated with stomatal control so as to maximize leaf photosynthesis. Drought and notching experiments were conducted on Pinus sylvestris, Picea abies, Betula Pendula and Populus tremula seedlings in glasshouse conditions to study the dependence of NSLs on leaf osmotic and water status, and their coordination with stomatal control, on timescales of minutes and weeks, to test the assumptions and predictions of the optimization hypothesis. Both NSLs and stomatal conductance followed power-law functions of leaf osmotic concentration and leaf water potential. Moreover, stomatal conductance was proportional to the square root of soil-to-leaf hydraulic conductance, as predicted by the optimization hypothesis. Though the detailed mechanisms underlying the dependence of NSLs on leaf osmotic or water status lie outside the scope of this study, our results support the hypothesis that NSLs and stomatal control are coordinated to maximize leaf photosynthesis and allow the effect of NSLs to be included in models of tree gas-exchange.

Soil fungi are a major factor maintaining plant diversity and productivity, but the underlying mechanisms are still poorly understood. Based on a biodiversity–ecosystem functioning experiment in southeast China, we evaluated the impacts of root-associated soil fungi on plant total, above- and belowground biomass production in monocultures and in different 2- and 4-tree species mixtures using two pools of four subtropical tree species each. All plots were inoculated with forest soil but half of the plots were additionally treated with fungicide to suppress fungi. Tree species richness promoted individual biomass only in control but not in fungicide-treated soils, leading to positive relative yield totals (RYTs, sum of the relative growth of each tree species in mixture to monoculture) of mixed species plots in control soils. Additive partitioning analysis showed that the net biodiversity effects were due to positive complementarity effects (CE) rather than selection effects. The relative yield of individual tree species was positively correlated with arbuscular mycorrhizal (AM) colonization for AM trees but not significantly correlated with ectomycorrhizal (EM) colonization for EM trees. The molecular analysis of the root-associated fungi showed that larger RYT and CE values were correlated with greater dissimilarity in pathogenic fungal communities between tree species, suggesting distinct pathogen compositions will result in overyielding and high complementarity in mixture.

Summary Experimental evidence that nonstomatal limitations to photosynthesis (NSLs) correlate with leaf sugar and/or leaf water status suggests the possibility that stomata adjust to maximise photosynthesis through a trade-off between leaf CO2 supply and NSLs, potentially involving source-sink interactions. However, the mechanisms regulating NSLs and sink strength, as well as their implications for stomatal control, remain uncertain. We used an analytically solvable model to explore optimal stomatal control under alternative hypotheses for source and sink regulation. We assumed that either leaf sugar concentration or leaf water potential regulates NSLs, and that either phloem turgor pressure or phloem sugar concentration regulates sink phloem unloading. All hypotheses lead to realistic stomatal responses to light, CO2 and air humidity, including conservative behaviour for the intercellular-to-atmospheric CO2 concentration ratio. Sugar- and water-regulated NSLs are distinguished by the presence/absence of a stomatal closure response to changing sink strength. Turgor- and sugar-regulated phloem unloading are distinguished by the presence/absence of stomatal closure under drought and avoidance/occurrence of negative phloem turgor. Results from girdling and drought experiments on Pinus sylvestris, Betula pendula, Populus tremula and Picea abies saplings are consistent with optimal stomatal control under sugar-regulated NSLs and turgor-regulated unloading. Our analytical results provide a simple representation of stomatal responses to above- and below-ground environmental factors and sink activity. Experimental evidence that nonstomatal limitations to photosynthesis (NSLs) correlate with leaf sugar and/or leaf water status suggests the possibility that stomata adjust to maximise photosynthesis through a trade-off between leaf CO2 supply and NSLs, potentially involving source-sink interactions. However, the mechanisms regulating NSLs and sink strength, as well as their implications for stomatal control, remain uncertain. We used an analytically solvable model to explore optimal stomatal control under alternative hypotheses for source and sink regulation. We assumed that either leaf sugar concentration or leaf water potential regulates NSLs, and that either phloem turgor pressure or phloem sugar concentration regulates sink phloem unloading. All hypotheses led to realistic stomatal responses to light, CO2 and air humidity, including conservative behaviour for the intercellular-to-atmospheric CO2 concentration ratio. Sugar-regulated and water-regulated NSLs are distinguished by the presence/absence of a stomatal closure response to changing sink strength. Turgor-regulated and sugar-regulated phloem unloading are distinguished by the presence/absence of stomatal closure under drought and avoidance/occurrence of negative phloem turgor. Results from girdling and drought experiments on Pinus sylvestris, Betula pendula, Populus tremula and Picea abies saplings are consistent with optimal stomatal control under sugar-regulated NSLs and turgor-regulated unloading. Our analytical results provide a simple representation of stomatal responses to above-ground and below-ground environmental factors and sink activity.

Mangroves often allocate a relatively large proportion of their total biomass production to their roots, and the belowground biomass of these forests contributes towards globally significant carbon sinks. However, little information is available on root production in mangroves due to the difficulties in carrying out measurements of belowground processes, particularly if there is regular flooding. In this study, we examined fine and coarse root production in the east coast of the Malaysian Peninsula. Ingrowth cores were used over the course of 17 months. In September 2014, twenty cores were randomly placed in each of five plots. Three cores were collected from each plot (fifteen cores in total), once every three months. Each core was divided into five 10 cm layers and root dry mass was recorded. Standing root biomass was also measured at the time of final collection using an additional 15 cores. There was a seasonal pattern in root production, which peaked in March and December 2015, after and during the monsoon season. Root biomass in the cores peaked at 33.23 ± 6.3 t ha−1 and 21.46 ± 7.3 t ha−1 in March and December respectively. Standing root biomass in February 2016 in the forest was 20.81 ± 2.8 t ha−1. After 17 months, the final root biomass in the cores was 14% less than the standing root biomass. These data suggest surprisingly rapid growth rates and turnover for mangrove roots. Total root biomass significantly increased with root depth and 78% of the roots, in all soil layers, consisted of fine roots (

Future climate will be characterized by an increase in frequency and duration of drought and warming that exacerbates atmospheric evaporative demand. How trees acclimate to long-term soil moisture changes and whether these long-term changes alter trees' sensitivity to short-term (day to months) variations of vapor pressure deficit (VPD) and soil moisture is largely unknown. Leaf gas exchange measurements were performed within a long-term (17 years) irrigation experiment in a drought-prone Scots pine-dominated forest in one of Switzerland's driest areas on trees in naturally dry (control), irrigated, and 'irrigation-stop' (after 11 years of irrigation) conditions. Seventeen years of irrigation increased photosynthesis (A) and stomatal conductance (g(s)) and reduced g(s) sensitivity to increasing VPD and soil drying. Following irrigation-stop, gas exchange decreased only after 3 years. After 5 years, maximum carboxylation (V-cmax) and electron transport (J(max)) rates in irrigation-stop recovered to similar levels as to before the irrigation-stop. These results suggest that long-term release from soil drought reduces the sensitivity to VPD and that atmospheric constraints may play an increasingly important role in combination with soil drought. Moreover, our study indicates that structural adjustments lead to an attenuation of initially strong leaf-level acclimation to strong multiple-year drought. Acclimation to irrigation increased gas exchange in Pinus sylvestris, but reduced the sensitivity to short-term changes. In addition, structural adjustments led to an attenuation of initially strong leaf-level acclimation.

Climate change will have a considerable impact on forest productivity worldwide. Forecasting the magnitude of such impact, with multiple environmental stressors changing simultaneously, is only possible with the help of process-based models. In order to assess their performance, such models require careful evaluation against measurements. However, direct comparison of model outputs against observational data is often not reliable, as models may provide the right answers due to the wrong reasons. This would severely hinder forecasting abilities under unprecedented climate conditions. Here, we present a methodology for model assessment, which supplements the traditional output-to-observation model validation. It evaluates model performance through its ability to reproduce observed seasonal changes of the most limiting environmental driver (MLED) for a given process, here daily gross primary productivity (GPP). We analyzed seasonal changes of the MLED for GPP in two contrasting pine forests, the Mediterranean Pinus halepensis Mill. Yatir (Israel) and the boreal Pinus sylvestris L. Hyytiälä (Finland) from three years of eddy-covariance flux data. Then, we simulated the same period with a state-of-the-art process-based simulation model (LandscapeDNDC). Finally, we assessed if the model was able to reproduce both GPP observations and MLED seasonality. We found that the model reproduced the seasonality of GPP in both stands, but it was slightly overestimated without site-specific fine-tuning. Interestingly, although LandscapeDNDC properly captured the main MLED in Hyytiälä (temperature) and in Yatir (soil water availability), it failed to reproduce high-temperature and high-vapor pressure limitations of GPP in Yatir during spring and summer. We deduced that the most likely reason for this divergence is an incomplete description of stomatal behavior. In summary, this study validates the MLED approach as a model evaluation tool, and opens up new possibilities for model improvement.

Stomatal conductance formulations are of great importance to how land surface models predict carbon assimilation and transpiration in vegetation. In this study, novel stomatal conductance formulations based on the CAP optimisation hypothesis (Dewar et al. 2018) are implemented in the land surface model JSBACH. Besides new stomatal conductance functions, the CAP framework enables a computational streamlining of the resolution of photosynthesis rate and leaf internal CO2 concentration.The formulations are based on the CAP optimisation hypothesis coupled to different photosynthesis models. Models constructed this way incorporate non-stomatal limitations to photosynthesis through the coupling of carbon assimilation to the soil-to-leaf hydraulic pathway. This entails a direct link from soil water status to stomatal conductance, photosynthesis rate and leaf internal CO2 concentration. While this construction does away with the need for some previous fitted or empirical parameters, new parameters are required to represent xylem hydraulic conductance and downregulation of photosynthesis during drought stress.These new models are compared to the widely used USO stomatal conductance model (Medlyn et al. 2011). A standalone version of JSBACH is run for single grid cells representing two boreal Scots pine (Pinus sylvestris) dominated sites in Finland (Hyytiälä and Sodankylä). Climate forcing is done with FLUXNET data from 2001 through 2012 and observations are from eddy covariance measurements from the two sites.Preliminary results indicate that some of the new formulations give reasonable results. This is very promising, since they are more detailed and theoretically robust than their semi-empirical predecessors, yet streamline the computational process.References:Dewar et al. 2018, New Phytol. 217: 571–581Medlyn et al. 2011, Glob. Change Biol. 17: 2134–2144

SummaryThe future climate will be characterized by an increase in frequency and duration of drought and warming that exacerbates atmospheric evaporative demand. How trees acclimate to long-term soil moisture changes and whether these long-term changes alter trees’ sensitivity to short-term (day to months) variations of vapor pressure deficit (VPD) and soil moisture is largely unknown.Leaf gas exchange measurements were performed within a long-term (17 years) irrigation experiment in a Scots pine-dominated forest in one of Switzerland’s driest areas on trees in naturally dry (control), irrigated, and‘irrigation-stop’ (after 11 years of irrigation) conditions.Seventeen years of irrigation increased photosynthesis (A) and stomatal conductance (gs) and reduced the gs sensitivity to increasing VPD but not to soil drying. Following irrigation-stop, gas exchange did not decrease immediately, but after three years, had decreased significantly in irrigation-stop trees. Vcmax and Jmax recovered after five years.These results suggest that long-term release of soil drought reduces the sensitivity to atmospheric evaporative demand and that atmospheric constraints may play an increasingly important role in combination with soil drought. In addition, they suggest that structural adjustments lead to an attenuation of initially strong leaf-level acclimation to strong multiple-year drought.

On-going climate change is increasing the risk of drought stress across large areas worldwide. Such drought events decrease ecosystem productivity and have been increasingly linked to tree mortality. Understanding how trees respond to water shortage is key to predicting the future of ecosystem functions. Phloem is at the core of the tree functions, moving resources such as non-structural carbohydrates, nutrients, and defence and information molecules across the whole plant. Phloem function and ability to transport resources is tightly controlled by the balance of carbon and water fluxes within the tree. As such, drought is expected to impact phloem function by decreasing the amount of available water and new photoassimilates. Yet, the effect of drought on the phloem has received surprisingly little attention in the last decades. Here we review existing knowledge on drought impacts on phloem transport from loading and unloading processes at cellular level to possible effects on long-distance transport and consequences to ecosystems via ecophysiological feedbacks. We also point to new research frontiers that need to be explored to improve our understanding of phloem function under drought. In particular, we show how phloem transport is affected differently by increasing drought intensity, from no response to a slowdown, and explore how severe drought might actually disrupt the phloem transport enough to threaten tree survival. Because transport of resources affects other organisms interacting with the tree, we also review the ecological consequences of phloem response to drought and especially predatory, mutualistic and competitive relations. Finally, as phloem is the main path for carbon from sources to sink, we show how drought can affect biogeochemical cycles through changes in phloem transport. Overall, existing knowledge is consistent with the hypotheses that phloem response to drought matters for understanding tree and ecosystem function. However, future research on a large range of species and ecosystems is urgently needed to gain a comprehensive understanding of the question.

Climate conditions in which tree species are able to grow are determined by their ecophysiological traits. The genus Pinus spp. is widespread across Eurasia, so that the different Pinus species have evolved to live within diverse climate envelops, from the boreal Scots pine (Pinus sylvestris L.) to the Mediterranean Aleppo pine (Pinus halepensis Mill.). Therefore, the different pine species are expected to present contrasting responses to environmental stressors, depending on the ones that populations had faced in the past.Here we analyze the impact of climate on stand carbon fluxes in two contrasting stands -i.e. a boreal Finnish Scots pine stand and a Mediterranean Israeli Aleppo pine stand. We use a machine learning approach -i.e. Random Forest algorithm- to evaluate seasonal changes in the most limiting environmental driver (MLED) for forest productivity. Then, we use data from controlled experiments with Aleppo and Scots pine saplings, in which we evaluated their response to drought and heat stresses, in order to assess if differences in their ecophysiological traits may explain their ability to grow in such contrasting climate conditions.Our results suggest that the MLED of forest productivity during all year in the boreal stand are low temperatures. Conversely but not surprisingly, the MLED in the Mediterranean stand is soil water availability, especially during summer. Therefore, we expect P. halepensis to be better adapted to heat and drought stresses, whereas we expect P. sylvestris to present higher photosynthetic rates at lower temperatures. Controlled experiments confirm these expectations, with a remarkable isohydric behavior of P. halepensis during drought, and different species responses of photosynthesis thermal optimum to heat and drought stress. Our results highlight the need to understand how traits determine tree species’ responses to different environmental stressors, in order to anticipate their performance in a warmer world.

Understanding stomatal regulation is fundamental to predicting the impact of changing environmental conditions on vegetation. However, the influence of soil temperature (ST) and soil water content (SWC) on canopy conductance (g(s)) through changes in belowground hydraulic conductance (k(bg)) remains poorly understood, because k(bg) has seldom been measured in field conditions. Our aim was to (a) examine the dependence of k(bg) on ST and SWC, (b) examine the dependence of g(s) on k(bg) and (c) test a recent stomatal optimization model according to which g(s) and soil-to-leaf hydraulic conductance are strongly coupled. We estimated k(bg) from continuous sap flow and xylem diameter measurements in three boreal species. k(bg) increased strongly with increasing ST when ST was below +8 degrees C, and typically increased with increasing SWC when ST was not limiting. g(s) was correlated with k(bg) in all three species, and modelled and measured g(s) were well correlated in Pinus sylvestris (a model comparison was only possible for this species). These results imply an important role for k(bg) in mediating linkages between the soil environment and leaf gas exchange. In particular, our finding that ST strongly influences k(bg) in mature trees may help us to better understand tree behaviour in cold environments.

Positive sap pressures are produced in the xylem of birch trees in boreal conditions during the time between the thawing of the soil and bud break. During this period, xylem embolisms accumulated during wintertime are refilled with water. The mechanism for xylem sap pressurization and its environmental drivers are not well known. We measured xylem sap flow, xylem sap pressure, xylem sap osmotic concentration, xylem and whole stem diameter changes, and stem and root non-structural carbohydrate concentrations, along with meteorological conditions at two sites in Finland during and after the sap pressurisation period. The diurnal dynamics of xylem sap pressure and sap flow during the sap pressurisation period varied, but were more often opposite to the diurnal pattern after bud burst, i.e. sap pressure increased and sap flow rate mostly decreased when temperature increased. Net conversion of soluble sugars to starch in the stem and roots occurred during the sap pressurisation period. Xylem sap osmotic pressure was small in comparison to total sap pressure, and it did not follow changes in environmental conditions or tree water relations. Based on these findings, we suggest that xylem sap pressurisation and embolism refilling occur gradually over a few weeks through water transfer from parenchyma cells to xylem vessels during daytime, and then the parenchyma are refilled mostly during nighttime by water uptake from soil. Possible drivers for water transfer from parenchyma cells to vessels are discussed. Also the functioning of thermal dissipation probes in conditions of changing stem water content is discussed.

Transpiration from the Amazon rainforest generates an essential water source at a global and local scale. However, changes in rainforest function with climate change can disrupt this process, causing significant reductions in precipitation across Amazonia, and potentially at a global scale. We report the only study of forest transpiration following a long-term (>10 year) experimental drought treatment in Amazonian forest. After 15 years of receiving half the normal rainfall, drought-related tree mortality caused total forest transpiration to decrease by 30%. However, the surviving droughted trees maintained or increased transpiration because of reduced competition for water and increased light availability, which is consistent with increased growth rates. Consequently, the amount of water supplied as rainfall reaching the soil and directly recycled as transpiration increased to 100%. This value was 25% greater than for adjacent non-droughted forest. If these drought conditions were accompanied by a modest increase in temperature (e.g. 1.5°C), water demand would exceed supply, making the forest more prone to increased tree mortality. This article is protected by copyright. All rights reserved.

Prerequisite for selection of appropriate tree species in afforestation programs is to understand their water use strategy. Acacia mangium Willd., Schima wallichii Choisy, and Cunninghamia lanceolata (Lamb.) Hook are the three main vegetation restoration pioneer species in southern China, but no comparative research on the water use strategy of these three tree species have been reported. Our objective was to gain a detailed understanding of how photosynthetically active radiation (PAR), vapor pressure deficit (VPD), and soil water content (SWC) at different soil depths control the sap flux density (Js) in the dry and wet seasons. We measured the Js of these three tree species by using the thermal dissipation method in low subtropical China. We found that both S. wallichii and C. lanceolata differed clearly in their stomatal behavior from one season to another, while A. mangium did not. The canopy conductance per sapwood area of S. wallichii and C. lanceolata was very sensitive to VPD in the dry season, but not in the wet season. The Js of A. mangium was negatively correlated to SWC in all soil layers and during both seasons, while the other two species were not sensitive to SWC in the deeper layers and only positively correlated to SWC in dry season. Our results demonstrate that the three species have distinct water use strategies and may therefore respond differently to changing climate.

Tree responses to altered water availability range from immediate (e.g. stomatal regulation) to delayed (e.g. crown size adjustment). The interplay of the different response times and processes, and their effects on long‐term whole‐tree performance, however, is hardly understood. - Here we investigated legacy effects on structures and functions of mature Scots pine in a dry inner‐Alpine Swiss valley after stopping an 11‐yr lasting irrigation treatment. Measured ecophysiological time series were analysed and interpreted with a system‐analytic tree model. - We found that the irrigation stop led to a cascade of downregulations of physiological and morphological processes with different response times. Biophysical processes responded within days, whereas needle and shoot lengths, crown transparency, and radial stem growth reached control levels after up to 4 yr only. Modelling suggested that organ and carbon reserve turnover rates play a key role for a tree’s responsiveness to environmental changes. Needle turnover rate was found to be most important to accurately model stem growth dynamics. - We conclude that leaf area and its adjustment time to new conditions is the main determinant for radial stem growth of pine trees as the transpiring area needs to be supported by a proportional amount of sapwood, despite the growth‐inhibiting environmental conditions. ISSN:0028-646X ISSN:1469-8137

Forests are sensitive to droughts, which increase the mortality rate of tree species. Various processes have been proposed to underlie drought-induced tree mortality, including hydraulic failure, carbon starvation and increased susceptibility to natural enemies. To give insights into these processes, we assessed the metabolic effects of a mortality-inducing drought on seedlings of Pinus sylvestris L. (Scots Pine), a widespread and important Eurasian species. We found divergence over time in the foliar metabolic composition of droughted vs well-watered seedlings, with the former showing increased abundance of aromatic amino acids and decreases in secondary metabolism associated with defence. We observed no significant differences amongst provenances in these effects: seedlings from drought-prone areas showed the same foliar metabolic changes under drought as seedlings from moist environments, although morphological effects of drought varied by provenance. Overall, our results demonstrate how severe drought prior to death may target particular primary and secondary metabolic pathways, weakening defences against natural enemies and contributing to the risk of drought-induced mortality in P. sylvestris.

Sugar transport in the phloem is driven by turgor pressure gradients which are created by osmotic gradients resulting from sugars loaded to the phloem at the source tissue and unloaded at the sink tissue. Therefore, osmolality is a key parameter that can be used to evaluate sugar status and get an indication of the driving force for phloem transport. Here we describe how osmotic concentration measurements from inner bark (practically, the phloem) and needles of trees can be measured. This protocol presents the procedure used by Lintunen et al. (Front Plant Sci 7:726, 2016) and Paljakka et al. (Plant Cell Environ 40:2160-2173, 2017), extended by practical advice and discussion of potential errors and caveats. We describe how to implement this procedure for gymnosperm as well as angiosperm trees. This method uses mechanical sap extraction with a centrifuge from inner bark and leaf samples, which have gone through a deep freeze treatment and thawing. The osmotic potential of these samples is then analyzed with a freezing point or vapor pressure osmometer. The aim of these measurements is to study the spatial and temporal dynamics of phloem function.

Complementary resource use has often been claimed to explain positive effects of plant diversity on ecosystem functioning, but the underlying mechanisms of complementarity have rarely been directly quantified in forest systems. The aim of this study was to characterize spatial and temporal water uptake of subtropical tree saplings, to assess the role of interspecific interaction on water uptake patterns and to quantify species niche breadth and overlap. Our experiment comprised two deciduous ( Castanea henryi , Quercus serrata ) and two evergreen tree species ( Elaeocarpus decipiens , Schima superba ) that were each planted in monoculture and 4-species mixture. We used deuterium-enriched water to trace seasonal water uptake from 5 cm and 20 cm soil depth. Castanea exploited predominantly the upper soil (74% of overall tracer uptake across treatments) whereas Quercus (50%), Elaeocarpus (57%) and Schima (62%) tended to use both soil layers more equally. Species identity had an overall significant effect on isotopic enrichment in stem water. There was no effect of species richness: niche breadth and overlap of single species was not affected by interspecific interactions in mixtures. Niche overlap between deciduous species was lowest (74%) whereas the two evergreen species had similar water uptake patterns (91%). According to our results, interspecific competition did not alter water uptake patterns of the studied species during the early phase of forest establishment. Thus, soil water uptake complementarity could only occur through inherent (fundamental) specific differences in water uptake niches based on sapling specialization, while phenotypic adjustments to interspecific interaction or neighbor diversity are less important.

Sugar transport in the phloem is driven by turgor pressure gradients which are created by osmotic gradients resulting from sugars loaded to the phloem at the source tissue and unloaded at the sink tissue. Therefore, osmolality is a key parameter that can be used to evaluate sugar status and get an indication of the driving force for phloem transport. Here we describe how osmotic concentration measurements from inner bark (practically, the phloem) and needles of trees can be measured. This protocol presents the procedure used by Lintunen et al. (Front Plant Sci 7:726, 2016) and Paljakka et al. (Plant Cell Environ 40:2160-2173, 2017), extended by practical advice and discussion of potential errors and caveats. We describe how to implement this procedure for gymnosperm as well as angiosperm trees. This method uses mechanical sap extraction with a centrifuge from inner bark and leaf samples, which have gone through a deep freeze treatment and thawing. The osmotic potential of these samples is then analyzed with a freezing point or vapor pressure osmometer. The aim of these measurements is to study the spatial and temporal dynamics of phloem function.

Increasing biodiversity has been linked to higher primary productivity in terrestrial ecosystems. However, the underlying ecophysiological mechanisms remain poorly understood. We investigated the effects of surrounding species richness (monoculture, two‐ and four‐species mixtures) on the ecophysiology of Lithocarpus glaber seedlings in experimental plots in subtropical China. A natural rain event isotopically labelled both the water uptaken by the L. glaber seedlings and the carbon in new photoassimilates through changes of photosynthetic discrimination. We followed the labelled carbon (C) and oxygen (O) in the plant–soil–atmosphere continuum. We measured gas‐exchange variables (C assimilation, transpiration and above‐ and belowground respiration) and δ13C in leaf biomass, phloem, soil microbial biomass, leaf‐ and soil‐respired CO 2 as well as δ18O in leaf and xylem water. The 13C signal in phloem and respired CO 2 in L. glaber in monoculture lagged behind those in species mixture, showing a slower transport of new photoassimilates to and through the phloem in monoculture. Furthermore, leaf‐water 18O enrichment above the xylem water in L. glaber increased after the rain in lower diversity plots suggesting a lower ability to compensate for increased transpiration. Lithocarpus glaber in monoculture showed higher C assimilation rate and water‐use efficiency. However, these increased C resources did not translate in higher growth of L. glaber in monoculture suggesting the existence of larger nongrowth‐related C sinks in monoculture. These ecophysiological responses of L. glaber, in agreement with current understanding of phloem transport are consistent with a stronger competition for water resources in monoculture than in species mixtures. Therefore, increasing species diversity in the close vicinity of the studied plants appears to alleviate physiological stress induced by water competition and to counterbalance the negative effects of interspecific competition on assimilation rates for L. glaber by allowing a higher fraction of the C assimilated to be allocated to growth in species mixture than in monoculture.

International audience; nitori Accumulating evidence highlights increased mortality risks for trees during severe drought, particularly under warmer temperatures and increasing vapour pressure deficit (VPD). Resulting forest die-off events have severe consequences for ecosystem services, biophysical and biogeochemical land-atmosphere processes. Despite advances in monitoring, modelling and experimental studies of the causes and consequences of tree death from individual tree to ecosystem and global scale, a general mechanistic understanding and realistic predictions of drought mortality under future climate conditions are still lacking. We update a global tree mortality map and present a roadmap to a more holistic understanding of forest mortality across scales. We highlight priority research frontiers that promote: (1) new avenues for research on key tree ecophysiological responses to drought; (2) scaling from the tree/plot level to the ecosystem and region; (3) improvements of mortality risk predictions based on both empirical and mechanistic insights; and (4) a global mong network of forest mortality. In light of recent and anticipated large forest die-off events such a research agenda is timely and needed to achieve scientific understanding for realistic predictions of drought-induced tree mortality. The implementation of a sustainable network will require support by stakeholders and political authorities at the international level.

Tree transpiration depends on biotic and abiotic factors that might change in the future, including precipitation and soil moisture status. Although short-term sap flux responses to soil moisture and evaporative demand have been the subject of attention before, the relative sensitivity of sap flux to these two factors under long-term changes in soil moisture conditions has rarely been determined experimentally. We tested how long-term artificial change in soil moisture affects the sensitivity of tree-level sap flux to daily atmospheric vapor pressure deficit ( VPD) and soil moisture variations, and the generality of these effects across forest types and environments using four manipulative sites in mature forests. Exposure to relatively long-term (two to six years) soil moisture reduction decreases tree sap flux sensitivity to daily VPD and relative extractable water ( REW ) variations, leading to lower sap flux even under high soil moisture and optimal VPD . Inversely, trees subjected to long-term irrigation showed a significant increase in their sensitivity to daily VPD and REW , but only at the most water-limited site. The ratio between the relative change in soil moisture manipulation and the relative change in sap flux sensitivity to VPD and REW variations was similar across sites suggesting common adjustment mechanisms to long-term soil moisture status across environments for evergreen tree species. Overall, our results show that long-term changes in soil water availability, and subsequent adjustments to these novel conditions, could play a critical and increasingly important role in controlling forest water use in the future.

The pull of water from the soil to the leaves causes water in the transpiration stream to be under negative pressure decreasing the water potential below zero. The osmotic concentration also contributes to the decrease in leaf water potential but withmuch lesser extent. Thus, the surface tension force is approximately balanced by a force induced by negative water potential resulting in concavely curved water-air interfaces in leaves. The lowered water potential causes a reduction in the equilibrium water vapor pressure in internal (sub-stomatal/ intercellular) cavities in relation to that over water with the potential of zero, i.e., over the flat surface. The curved surface causes a reduction also in the equilibrium vapor pressure of dissolved CO2, thus enhancing its physical solubility to water. Although the water vapor reduction is acknowledged by plant physiologists its consequences for water vapor exchange at low water potential values have received very little attention. Consequences of the enhanced CO2 solubility to a leaf water-carbon budget have not been considered at all before this study. We use theoretical calculations and modeling to show how the reduction in the vapor pressures affects transpiration and carbon assimilation rates. Our results indicate that the reduction in vapor pressures of water and CO2 could enhance plant water use efficiency up to about 10% at a leaf water potential of -2 MPa, and much more when water potential decreases further. The low water potential allows for a direct stomatal water vapor uptake from the ambient air even at sub-100% relative humidity values. This alone could explain the observed rates of foliar water uptake by e.g., the coastal redwood in the fog belt region of coastal California provided the stomata are sufficiently open. The omission of the reduction in the water vapor pressure causes a bias in the estimates of the stomatal conductance and leaf internal CO2 concentration based on leaf gas exchange measurements. Manufactures of leaf gas exchange measurement systems should incorporate leaf water potentials in measurement set-ups.

The pull of water from the soil to the leaves causes water in the transpiration stream to be under negative pressure decreasing the water potential below zero. The osmotic concentration also contributes to the decrease in leaf water potential but with much lesser extent. Thus, the surface tension force is approximately balanced by a force induced by negative water potential resulting in concavely curved water-air interfaces in leaves. The lowered water potential causes a reduction in the equilibrium water vapor pressure in internal (sub-stomatal/intercellular) cavities in relation to that over water with the potential of zero, i.e., over the flat surface. The curved surface causes a reduction also in the equilibrium vapor pressure of dissolved CO2, thus enhancing its physical solubility to water. Although the water vapor reduction is acknowledged by plant physiologists its consequences for water vapor exchange at low water potential values have received very little attention. Consequences of the enhanced CO2 solubility to a leaf water-carbon budget have not been considered at all before this study. We use theoretical calculations and modeling to show how the reduction in the vapor pressures affects transpiration and carbon assimilation rates. Our results indicate that the reduction in vapor pressures of water and CO2 could enhance plant water use efficiency up to about 10% at a leaf water potential of −2 MPa, and much more when water potential decreases further. The low water potential allows for a direct stomatal water vapor uptake from the ambient air even at sub-100% relative humidity values. This alone could explain the observed rates of foliar water uptake by e.g., the coastal redwood in the fog belt region of coastal California provided the stomata are sufficiently open. The omission of the reduction in the water vapor pressure causes a bias in the estimates of the stomatal conductance and leaf internal CO2 concentration based on leaf gas exchange measurements. Manufactures of leaf gas exchange measurement systems should incorporate leaf water potentials in measurement set-ups.

Values (Δi) predicted by a simplified photosynthetic discrimination model, based only on diffusion through air followed by carboxylation, are often used to infer ecological conditions from the 13C signature of plant organs (δ13Cp). Recent studies showed that additional isotope discrimination (d that includes mesophyll conductance, photorespiration and day respiration, and post-carboxylation discrimination) can strongly affect δ13Cp; however, little is known about its variability during plant ontogeny for different species. Effect of ontogeny on leaf gas exchange rates, Δi, observed discrimination (Δp) and d in leaf, phloem and root of seven herbaceous species at three ontogenetic stages were investigated under controlled conditions. Functional group identity and ontogeny significantly affected Δi and Δp. However, predicted Δi did not match Δp. d, strongly affected by functional group identity and ontogeny, varied by up to 14‰. d scaled tightly with stomatal conductance, suggesting complex controls including changes in mesophyll conductance. The magnitude of the changes in δ13Cp due to ontogeny was similar to that due to environmental factors reported in other studies. d and ontogeny should, therefore, be considered in ecosystem studies, integrated in ecosystem models using δ13Cp and limit the applicability of δ13Cleaf as a proxy for water-use efficiency in herbaceous plants.

Starch and soluble sugars are the major photosynthetic products, and their carbon isotope signatures reflect external versus internal limitations of CO2 fixation. There has been recent renewed interest in the isotope composition of carbohydrates, mainly for use in CO2 flux partitioning studies at the ecosystem level. The major obstacle to the use of carbohydrates in such studies has been the lack of an acknowledged method to isolate starch and soluble sugars for isotopic measurements. We here report on the comparison and evaluation of existing methods (acid and enzymatic hydrolysis for starch; ion-exchange purification and compound-specific analysis for sugars). The selectivity and reproducibility of the methods were tested using three approaches: (i) an artificial leaf composed of a mixture of isotopically defined compounds, (ii) a C4 leaf spiked with C3 starch, and (iii) two natural plant samples (root, leaf). Starch preparation methods based on enzymatic or acid hydrolysis did not yield similar results and exhibited contaminations by non-starch compounds. The specificity of the acidic hydrolysis method was especially low, and we therefore suggest terming these preparations as HCl-hydrolysable carbon, rather than starch. Despite being more specific, enzyme-based methods to isolate starch also need to be further optimized to increase specificity. The analysis of sugars by ion-exchange methods (bulk preparations) was fast but produced more variable isotope compositions than compound-specific methods. Compound-specific approaches did not in all cases correctly reproduce the target values, mainly due to unsatisfactory separation of sugars and background contamination. Our study demonstrates that, despite their wide application, methods for the preparation of starch and soluble sugars for the analysis of carbon isotope composition are not (yet) reliable enough to be routinely applied and further research is urgently needed to resolve the identified problems. Copyright © 2009 John Wiley & Sons, Ltd.

Secondary compounds play multiple ecological roles. In this study, we present novel experimental evidence of differential tolerance to freezing temperatures among chemotypes of a chemically polymorphic plant, Thymus vulgaris. Non-phenolic chemotypes showed a significantly better survival and re-growth after early-winter freezing (−10° in early December) than phenolic chemotypes. Comparison of temperature data (1971–2002) at a phenolic and non-phenolic site showed that whereas early-winter freezing occurred in 6 years in the non-phenolic site they never occurred at the phenolic site. Observations of trichome morphology (where the essential oil is stocked) with and without intense freezing indicate that non-phenolic chemotypes may escape any negative effects of freezing by releasing their essential oil into the atmosphere during severe freezing. The correlation between tolerance of freezing and local temperature regimes strongly suggests that differential freezing resistance is a key ingredient of the distribution of thyme chemotypes in space.

Altres ajuts: F.M. acknowledges support from the STReSS COST action (FP1106) for short-term scientific missions to Edinburgh and Barcelona. M.M. and Y.S. acknowledge support from NERC (project NE/IO107749/1). Accurate modelling of drought-induced mortality is challenging. A steady-state model is presented integrating xylem and phloem transport, leaf-level gas exchange and plant carbohydrate consumption during drought development. A Bayesian analysis of parameter uncertainty based on expert knowledge and a literature review is carried out. The model is tested by combining six data compilations covering 170 species using information on sensitivities of xylem conductivity, stomatal conductance and leaf turgor to water potential. The possible modes of plant failure at steady state are identified (i.e. carbon (C) starvation, hydraulic failure and phloem transport failure). Carbon starvation occurs primarily in the parameter space of isohydric stomatal control, whereas hydraulic failure is prevalent in the space of xylem susceptibility to embolism. Relative to C starvation, phloem transport failure occurs under conditions of low sensitivity of photosynthesis and high sensitivity of growth to plant water status. These three failure modes are possible extremes along two axes of physiological vulnerabilities, one characterized by the balance of water supply and demand and the other by the balance between carbohydrate sources and sinks. Because the expression of physiological vulnerabilities is coordinated, we argue that different failure modes should occur with roughly equal likelihood, consistent with predictions using optimality theory.